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Vol. 15, Issue 2, 838-850, February 2004
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* CNRS Centre de Génétique Moléculaire, 91198 Gif-sur-Yvette, France;
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115; and
Department of Molecular, Cell and Developmental Biology, Sinsheimer Laboratories, University of California at Santa Cruz, Santa Cruz, California 95064
Submitted June 26, 2003;
Revised October 9, 2003;
Accepted October 10, 2003
Monitoring Editor: Mary Beckerle
The early Drosophila embryo undergoes two distinct membrane invagination events believed to be mechanistically related to cytokinesis: metaphase furrow formation and cellularization. Both involve actin cytoskeleton rearrangements, and both have myosin II at or near the forming furrow. Actin and myosin are thought to provide the force driving membrane invagination; however, membrane addition is also important. We have examined the role of myosin during these events in living embryos, with a fully functional myosin regulatory light-chain-GFP chimera. We find that furrow invagination during metaphase and cellularization occurs even when myosin activity has been experimentally perturbed. In contrast, the basal closure of the cellularization furrows and the first cytokinesis after cellularization are highly dependent on myosin. Strikingly, when ingression of the cellularization furrow is experimentally inhibited by colchicine treatment, basal closure still occurs at the appropriate time, suggesting that it is regulated independently of earlier cellularization events. We have also identified a previously unrecognized reservoir of particulate myosin that is recruited basally into the invaginating furrow in a microfilament-independent and microtubule-dependent manner. We suggest that cellularization can be divided into two distinct processes: furrow ingression, driven by microtubule mediated vesicle delivery, and basal closure, which is mediated by actin/myosin based constriction.
Abbreviations used: GFP, green fluorescent protein; Drok, Drosophila rho kinase; MHC, myosin heavy chain; MLCK; RLC, regulatory myosin light chain; TLCM, time-lapse confocal microscopy.
Online version of this article contains supplemental material and videos. Online version is available at www.molbiolcell.org.
Corresponding author. E-mail address: karess{at}cgm.cnrs-gif.fr.
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